Methods for expansion of human embryonic stem
Methods for expansion of human embryonic stem
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DOI：
10.1634/stemcells.2004-0297
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发表时间：
2005-05-01
期刊：
影响因子：
5.2
通讯作者：
Moon, SY
中科院分区：
文献类型：
--
作者：
Oh, SK;Kim, HS;Moon, SY
操纵人类胚胎干细胞(HESCs)需要精湛的技能。在这里，我们根据实验目的介绍了用于hESCs的机械转移和酶转移两种方法。我们采用机械交接的方法对hESC线路进行维护。尽管该方法费时费力，但该技术可以有效地转移未分化的hESCs，并产生类似的集落大小。当我们需要大量生产各种实验所需的细胞时，我们会实施酶转移法。酶处理的扩张在有限的时间内迅速产生更多的hESCs。然而，细胞团的大小不同，分化和未分化的细胞都有可能被转移。在有分化集落的情况下，两种方法的结合可以通过在酶处理之前通过人工选择从传代中排除分化集落来大量生产hESCs。
The manipulation of human embryonic stem cells (hESCs) requires refined skills. Here we introduce both mechanical and enzymatic transfer methods for hESCs depending on experimental purpose. We use the mechanical transfer method for maintenance of hESC lines. Although the method is laborious and time-consuming, the technique permits efficient transfer of undifferentiated hESCs and results in similar clump sizes. We implement the enzymatic transfer method when we need the bulk production of cells for various experiments. The enzyme-treated expansion rapidly produces greater amounts of hESCs within a limited time frame. However, the cell clumps vary in size, and there is a probability that both the differentiated and undifferentiated cells will be transferred,. In cases in which there are differentiated colonies, the combination of two methods allows mass production of hESCs by excluding differentiated colonies from passage by manual selection before enzyme treatment.